[unreadable] The overall objective of this proposal is to make real-time observations of gene expression in live cells at the transcription level and translation level. [unreadable] [unreadable] The specific aims of the proposed studies are: [unreadable] [unreadable] 1. To demonstrate the use of hybrid PNA beacons in order to detect sequence-specific m-RNA in live cells with high sensitivity and durability. [unreadable] [unreadable] 2. To develop new reporter proteins with short maturation time and short cellular lifetime in order to detect transient and stochastic behavior of gene expression of low copy number proteins. [unreadable] [unreadable] 3. To perfect and integrate state-of-the-art fluorescence microscopy with the hybrid PNA beacons and reporter genes in order to observe stochastic expression of a single gene at the transcriptional and translational levels. [unreadable] [unreadable] The proposed work, though highly exploratory in nature, will yield fundamental knowledge of gene expression, as well as general methodologies for biotechnology and clinical diagnostics. The new approach is likely to have a broad impact on biomedical research and lead to unexpected discoveries. [unreadable] [unreadable] [unreadable]